Direct Visualization of DNA Recognition by Restriction Endonuclease EcoRI

Chiung Fang Huang, Pei Wen Peng, Chih Ming Cheng, Jing Shin Tsai, Wei Ting Wang, Chien Ting Hsu, Keng Liang Ou, Tzu Sen Yang

研究成果: 雜誌貢獻文章

1 引文 (Scopus)

摘要

Background/Purpose: Restriction endonuclease is found in bacteria and can precisely cut off the DNA sequence specificity invaded from outside. Using the restriction endonuclease to cleave the double-stranded DNA at specific recognition nucleotide sequences is regarded as important biotechnology. However, traditional approaches cannot directly observe the process of specific binding of DNA by the restriction endonuclease. Methods: Single-molecule biotechnologies, including an optical tweezers system and single-molecule fluorescence detection system, were proposed to directly observe how the restriction endonuclease EcoRI found the recognition-binding site. Results: Overall, the study showed that EcoRI conjugated quantum dots (QDs) possessed the ability to recognize its specific binding sites. In addition, the distributions of the QD-EcoRI trajectory in the transverse direction were nearly the same; however, the difference occurred in the longitudinal direction along the DNA axis during the early and late stages of EcoRI recognition. Conclusion: These results suggest one-dimensional sliding of EcoRI existed along the DNA contour during EcoRI recognition. In addition, when EcoRI slided along the DNA contour, it would require more than 400 seconds for EcoRI to correctly find its DNA target site.
原文英語
頁(從 - 到)25-29
頁數5
期刊Journal of Experimental and Clinical Medicine(Taiwan)
5
發行號1
DOIs
出版狀態已發佈 - 2013

指紋

Deoxyribonuclease EcoRI
DNA Restriction Enzymes
DNA
Quantum Dots
Biotechnology
Binding Sites
Optical Tweezers
Optical Devices
Deoxyribonuclease I
Fluorescence
Bacteria

ASJC Scopus subject areas

  • Medicine(all)

引用此文

Direct Visualization of DNA Recognition by Restriction Endonuclease EcoRI. / Huang, Chiung Fang; Peng, Pei Wen; Cheng, Chih Ming; Tsai, Jing Shin; Wang, Wei Ting; Hsu, Chien Ting; Ou, Keng Liang; Yang, Tzu Sen.

於: Journal of Experimental and Clinical Medicine(Taiwan), 卷 5, 編號 1, 2013, p. 25-29.

研究成果: 雜誌貢獻文章

Huang, Chiung Fang ; Peng, Pei Wen ; Cheng, Chih Ming ; Tsai, Jing Shin ; Wang, Wei Ting ; Hsu, Chien Ting ; Ou, Keng Liang ; Yang, Tzu Sen. / Direct Visualization of DNA Recognition by Restriction Endonuclease EcoRI. 於: Journal of Experimental and Clinical Medicine(Taiwan). 2013 ; 卷 5, 編號 1. 頁 25-29.
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abstract = "Background/Purpose: Restriction endonuclease is found in bacteria and can precisely cut off the DNA sequence specificity invaded from outside. Using the restriction endonuclease to cleave the double-stranded DNA at specific recognition nucleotide sequences is regarded as important biotechnology. However, traditional approaches cannot directly observe the process of specific binding of DNA by the restriction endonuclease. Methods: Single-molecule biotechnologies, including an optical tweezers system and single-molecule fluorescence detection system, were proposed to directly observe how the restriction endonuclease EcoRI found the recognition-binding site. Results: Overall, the study showed that EcoRI conjugated quantum dots (QDs) possessed the ability to recognize its specific binding sites. In addition, the distributions of the QD-EcoRI trajectory in the transverse direction were nearly the same; however, the difference occurred in the longitudinal direction along the DNA axis during the early and late stages of EcoRI recognition. Conclusion: These results suggest one-dimensional sliding of EcoRI existed along the DNA contour during EcoRI recognition. In addition, when EcoRI slided along the DNA contour, it would require more than 400 seconds for EcoRI to correctly find its DNA target site.",
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AU - Hsu, Chien Ting

AU - Ou, Keng Liang

AU - Yang, Tzu Sen

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AB - Background/Purpose: Restriction endonuclease is found in bacteria and can precisely cut off the DNA sequence specificity invaded from outside. Using the restriction endonuclease to cleave the double-stranded DNA at specific recognition nucleotide sequences is regarded as important biotechnology. However, traditional approaches cannot directly observe the process of specific binding of DNA by the restriction endonuclease. Methods: Single-molecule biotechnologies, including an optical tweezers system and single-molecule fluorescence detection system, were proposed to directly observe how the restriction endonuclease EcoRI found the recognition-binding site. Results: Overall, the study showed that EcoRI conjugated quantum dots (QDs) possessed the ability to recognize its specific binding sites. In addition, the distributions of the QD-EcoRI trajectory in the transverse direction were nearly the same; however, the difference occurred in the longitudinal direction along the DNA axis during the early and late stages of EcoRI recognition. Conclusion: These results suggest one-dimensional sliding of EcoRI existed along the DNA contour during EcoRI recognition. In addition, when EcoRI slided along the DNA contour, it would require more than 400 seconds for EcoRI to correctly find its DNA target site.

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